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Journal: Clinical cancer research : an official journal of the American Association for Cancer Research
Article Title: Targeting the DNA Damage Response Pathways and Replication Stress in Colorectal Cancer
doi: 10.1158/1078-0432.CCR-22-0875
Figure Lengend Snippet: The proposed preclinical and clinical flow to potentially predict ATRi-sensitive and ATRi-resistant colorectal cancer tumors. After written consent of the patient, tumor sample can be either processed as FFPE sample for direct immunohistologic and immunofluorescence analysis, or preclinical models for in vitro and in vivo analyses can be established. Samples can be tested for direct in vitro drug screenings or for biomarkers analysis through immunofluorescence or IHC assays. To evaluate the relevance of the “composite biomarker” of sensitivity to ATR inhibition, we propose to detect the expression level of proteins ATM, RAD51, and RAD51C together with scoring of phospho-RPA32 at basal level—prior to treatment. Also, scoring of activated DNA-PK and RAD51 upon treatment with ATRi will be informative. This information may eventually lead to the identification of patients who might benefit from ATR inhibition monotherapy, and directly translate the knowledge from bench to bedside. FFPE, formalin-fixed, paraffin-embedded; PDO, patient-derived organoid; PDX, patient-derived xenograft; TTT, treatment. This figure was created with biorender.com.
Article Snippet: Organoids were then incubated with 1% BSA in PBS for 60 minutes, followed by incubation overnight with the following primary antibodies diluted in PBS containing 1% of BSA and 1% of donkey serum: anti-RAD51 (Millipore ABE257; 1:100),
Techniques: Immunofluorescence, In Vitro, In Vivo, Biomarker Assay, Inhibition, Expressing, Formalin-fixed Paraffin-Embedded, Derivative Assay